Current revision

Purpose

Procedure

Lysozyme in powder form from chicken egg white obtained from Signma® is used to make the lysozyme stock solution.

5mL of 70μM lysozyme stock solution was made in the following way:

0.005g of lysozyme was weighted out

0.005g of lysozyme was transferred to a 15ml falcon tube and mixed well with 5mL of distilled water.

See Melissa's Notebook for calculation that stated that 0.005g of lysozyme was needed for 5mL of 70μM of lysozyme stock solution.

The HAuCl4 used to make Au/Lysozyme samples was already made on 2012/09/05, and the concentration of the HAuCl4 was 10mM.

Au/Lysozyme samples was made with mole ratios listed below:

20 - 30 - 40 - 50 - 60 - 70 - 80 - 100 - 120 - 130

Based off the calculations for Au/BSA samples, a few changes were made to calculate the amount of lysozyme and HAuCl4 needed for Au/Lysozyme samples.

The table below listed the initial conditions for both lysozyme and HAuCl4 both theoretically and actually:

-

Lysozyme

HAuCl4

Molecular Weight [g/mol]

14300

339.79

Theoretical Concentration [M]

7.00E-05

0.01

Volume of Water to add [L]

0.005

0.025

Moles Present [mol]

3.50E-07

0.00025

Theoretical Mass Needed [g]

0.005

0.0849

Actual Mass Weighted [g]

0.005

0.0892

Actual Concentration [M]

0.0007

10501

With the above calculation, the actual concentration of lysozyme came out to be 70μM and the actual concentration of HAuCl4 came out to be 10.501mM. The two concentration was used to calculate the amount of water, lysozyme, HAuCl4 needed to make the ratios listed above.

The table below listed the amount of water, lysozyme, HAuCl4 used for each Au/Lysozyme sample:

Au/Lysozyme ratio

Lysozyme added [uL]

HAuCl4 added [uL]

Water added [uL]

Final Concentration of Au[mM]

Final Concentration of Lysozyme[uM]

20

114.3

15.2

7870.5

1

20

30

114.3

22.9

7862.8

1

30

40

114.3

30.5

7855.2

1

40

50

114.3

38.1

7847.6

1

50

60

114.3

45.7

7840

1

60

70

114.3

53.3

7832.4

1

70

80

114.3

60.9

7824.8

1

80

100

114.3

76.2

7809.5

1

100

120

114.3

91.4

7794.3

1

120

130

114.3

99

7786.7

1

130

The stock solutions were added with pipetman and non-sterile pipette tips. All samples were made in 10mL large test tubes. All samples were capped and wrapped in aluminum foil.

All samples were placed in incubator at 85°C for four hours then in room temperature to cool down.